摘要
【目的】研究不同浓度大蒜素对hPDLCsⅠ型胶原合成的影响及对hPDLF培养上清液胶原分泌的影响,探索大蒜素对hPDLCs胶原代谢的影响。
【材料及方法】本实验采用组织块培养法建立hPDLCs有限细胞系并绘制生长曲线。分别用(3、6、9ug/ml)大蒜素作用于第5代hPDLCs。用细胞免疫荧光化学方法标记Ⅰ型胶原,在荧光显微镜下观察并测得荧光值。化学比色法测定细胞培养上清液中羟脯氨酸含量。
【结果】使用组织块培养法进行hPDLCs的原代培养成功培养出了hPDLCs,经免疫组化确定。细胞生长曲线接近S形,符合一般细胞生长的规律。(3、6、9ug/ml)三种浓度大蒜素组均可使细胞上清液中羟脯氨酸含量显著降低,据此计算出的胶原蛋白含量,与对照组相比差异有显著性(p＜0.05),并具有剂量效应关系。(3、6、9ug/ml)三种浓度大蒜素可使hPDLCs胞浆内Ⅰ型胶原合成减少,与对照组相比差异有显著性(p＜0.05),并具有剂量效应关系。
【结论】1.大蒜素在一定的浓度范围内可抑制hPDLCs胶原分泌。2.大蒜素在一定的浓度范围内可抑制hPDLCsⅠ型胶原合成。
[Objectives] : The purpose of this study was to analyse the effects of Garlicin of different doses on human periodontal ligament cells collagen I synthesis and collagen secretion, providing more evidence for the clinical use of Garlicin in stomatology.
[Materials and Methods]: The hPDLCs were obtained from normal human ligament and cultured in vitro. The hPDLCs were exposed in culture to different Garlicin doses (3、6、9ug/ml) respectively for 48 hours.After that , intracellular expressin of collagen I were detected by Immunofluorescent microscopy,and the hydroxyproline in cell culture medium was determined by hydrolysismethod.
[Results] : The hPDLCs was cultured successfully,and the immunohistochemical studies demonstrated that the growing cells were fibroblasts and originated from mesenchymal.Garlicin could inhibit collagen I synthesis in hPDLCs from 3ug/ml to 9ug/ml.Garlicin also reduced hydroxyproline in cell culture medium in 3ug/ml、6ug/ml、9ug/ml group, and have the dose-effect relationship.
[Conclusions] : Garlicin chould depress hPDLCs collagen I synthesis and collagen excete in a certain dose range.The further use of Garlicin in Endontics needs more research.
引文
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